Subtypes of non-transformed human mammary epithelial cells cultured in vitro: histo-blood group antigen H type 2 defines basal cell-derived cells

Abstract. Normal (non-transformed) human mammary epithelial cell lines derived from reduction mammoplasties were analyzed by immunocytochemistry with more than 80 monoclonal antibodies (mAbs) and other specific reagents to tissue-specific and developmentally regulated antigens at different passage levels. A subpopulation of poorly differentiated, proliferating epithelial cells, corresponding to the 'selected' cell type of late passages, is shown to be characterized by a new marker, the histo-blood group antigen H type 2, probably carried on a membrane-bound glycolipid. These cells also express a number of other onco-developmental carbohydrate antigens [Le^y, Le^x, sialosyl-Le^a, precursor of Thomsen Friedenreich antigen (T_n), but not Thomsen-Friedenreich antigen and sialosyl-T_n]. Their cytokeratin (CK) phenotype, as assessed by reactivity with monospecific mAbs and two-dimensional gel electrophoresis, is CK 5, 6, 14 and 17, with CK 19 being consistently absent, and varying minor amounts of CK 7, 8 and 18, as well as 15 and 16. The reactivity of these cells with a panel of 11 mAbs specific for CK 18 varies considerably even after cloning, indicating heterogeneity of epitope expression or accessibility. Our data strongly suggest that the H type 2⁺ cells develop from the basal cell layer of the mammary gland.     

Limited Intermixing of Synaptic Vesicle Components upon Vesicle Recycling

Synaptic vesicles recycle repeatedly in order to maintain synaptic transmission. We have previously proposed that upon exocytosis the vesicle components persist as clusters, which would be endocytosed as whole units. It has also been proposed that the vesicle components diffuse into the plasma membrane and are then randomly gathered into new vesicles. We found here that while strong stimulation (releasing the entire recycling pool) causes the diffusion of the vesicle marker synaptotagmin out of synaptic boutons, moderate stimulation (releasing ～19% of all vesicles) is followed by no measurable diffusion. In agreement with this observation, synaptotagmin molecules labeled with different fluorescently tagged antibodies did not appear to mix upon vesicle recycling, when investigated by subdiffraction resolution stimulated emission depletion (STED) microscopy. Finally, as protein diffusion from vesicles has been mainly observed using molecules tagged with pH‐sensitive green fluorescent protein (pHluorin), we have also investigated the membrane patterning of several native and pHluorin‐tagged proteins. While the native proteins had a clustered distribution, the GFP‐tagged ones were diffused in the plasma membrane. We conclude that synaptic vesicle components intermix little, at least under moderate stimulation, possibly because of the formation of clusters in the plasma membrane. We suggest that several pHluorin‐tagged vesicle proteins are less well integrated in clusters.     

Subtypes of non-transformed human mammary epithelial cells cultured in vitro: histo-blood group antigen H type 2 defines basal cell-derived cells

Abstract. Normal (non-transformed) human mammary epithelial cell lines derived from reduction mammoplasties were analyzed by immunocytochemistry with more than 80 monoclonal antibodies (mAbs) and other specific reagents to tissue-specific and developmentally regulated antigens at different passage levels. A subpopulation of poorly differentiated, proliferating epithelial cells, corresponding to the 'selected' cell type of late passages, is shown to be characterized by a new marker, the histo-blood group antigen H type 2, probably carried on a membrane-bound glycolipid. These cells also express a number of other onco-developmental carbohydrate antigens [Le^y, Le^x, sialosyl-Le^a, precursor of Thomsen Friedenreich antigen (T_n), but not Thomsen-Friedenreich antigen and sialosyl-T_n). Their cytokeratin (CK) phenotype, as assessed by reactivity with monospecific mAbs and two-dimensional gel electrophoresis, is CK 5, 6, 14 and 17, with CK 19 being consistently absent, and varying minor amounts of CK 7, 8 and 18, as well as 15 and 16. The reactivity of these cells with a panel of 11 mAbs specific for CK 18 varies considerably even after cloning, indicating heterogeneity of epitope expression or accessibility. Our data strongly suggest that the H type 2⁺ cells develop from the basal cell layer of the mammary gland.     

Über die Entwicklung der Schuppenorgane und der Genitalanhänge in Abhängigkeit vom Hormonsystem beiLepisma saccharina L.

Zusammenfassung Die Entstehung der Schuppenorgane vonLepisma verläuft in ihren Grundzügen wie bei den Lepidopteren: Eine Schuppenstammzelle wird bei einer ersten differentiellen Teilung, bei der die Spindelachse senkrecht zur Körperoberfläche in der Epidermis steht, in zwei Tochterzellen geteilt. Eine von ihnen wird in einer zweiten differentiellen Teilung, deren Spindelachse schräg zur Körperoberfläche in der Epidermis liegt, in eine Schuppen- und eine Balgbildungszelle geteilt. Der zweite aus der senkrechten Mitose hervorgegangene Kern degeneriert aber nicht wie der ihm entsprechende Kern bei den Schuppenorganen der Lepidopteren, er wird auch nicht zu einem Sinneszellkern wie bei Borstenorganen von Lepidopteren oder zu einem Nebenzellkern des Schuppenorgans.Die Spindelachsen der schrägen Mitosen stehen in keiner festen Beziehung zur Körperlängsachse, wie das bei den Lepidopteren der Fall ist. Während der Schuppenbildungsphase sind Schuppen- und Balgkerne jedoch in Richtung der Körperlängsachse orientiert. Die Anordnung kommt dadurch zustande, daß die Balgkerne sich ausrichten.Bei Lepisma besteht eine Korrelation zwischen Körpergröße und Länge der Genitalanhänge, in der Weise, daß eine bestimmte Entwicklungsstufe der Geschlechtsanhänge eine bestimmte Mindestgröße der Tiere voraussetzt. Die höchstmögliche Entwicklungsstufe ist aber nicht bei allen Tieren dieser Körpergröße erreicht.Werden die Corpora allata zerstört, so bleibt der Zusammenhang zwischen der Entwicklungsstufe der Genitalanhänge und der Körpergröße bestehen; es können nicht etwa wie bei anderen Insekten beliebig kleine Tiere zu Imagines werden. Das Fehlen der Corpora allata wirkt sich in der Weise aus, daß mehr Tiere als normalerweise den bei einer bestimmten Körpergröße höchstmöglichen Entwicklungszustand erreichen.Die Arbeit wurde von Herrn Professor Dr. H.Piepho angeregt und von der Deutschen Forschungsgemeinschaft unterstützt.